hh Human Nephrin Antibody AF4269: R&D Systems

Human Nephrin Antibody

(2 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Nephrin in direct ELISAs and Western blots. In direct ELISAs, less than 20% cross-reactivity with recombinant mouse Nephrin is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Nephrin
    Gln23-Thr1029
    Accession # O60500
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunohistochemistry
    1-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunohistochemistry
Nephrin in Human Kidney. Nephrin was detected in immersion fixed paraffin-embedded sections of human kidney using 1.7 µg/mL Sheep Anti-Human Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4269) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to podocytes in glomeruli. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Nephrin by Western Blot. Western blot shows lysates of human kidney tissue under reducing and non-reducing conditions. PVDF membrane was probed with 1 µg/mL Sheep Anti-Human Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4269) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band for Nephrin was detected at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Nephrin
Nephrin, also known as renal glomerulus-specific cell adhesion receptor and nephrosis 1 (NPHS1) is a 185-200 kDa type I transmembrane protein belonging to the immunoglobulin (Ig) superfamily. It is expressed on podocytes and is an essential component of the interpodocyte-spanning slit diaphragm complex. Nephrin forms cis-hetero-oligomeric complexes with Neph1, followed by trans-homophilic interaction with Nephrin on opposing cells. Mutations in the Nephrin gene is the pathogenic cause of congenital nephrotic syndrome. Mature human Nephrin contains a 1033 aa extracellular region and a 165 aa cytoplasmic tail. One potential soluble splice form is known where aa’s 1056-1095 are deleted, eliminating the transmembrane region. Over aa 23-1029, human Nephrin shares 84% and 89% aa sequence identity with mouse and canine Nephrin, respectively.
  • Entrez Gene IDs:
    4868 (Human); 54631 (Mouse); 64563 (Rat)
  • Alternate Names:
    CNF; Nephrin; nephrosis 1, congenital, Finnish type (nephrin); NPHNCNF; NPHS1; Renal glomerulus-specific cell adhesion receptor
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. ELMO1 protects renal structure and ultrafiltration in kidney development and under diabetic conditions
    Sci Rep, 2016;6(0):37172.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC - Paraffin embedded
  2. Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis.
    Authors: Takasato M, Er P, Chiu H, Maier B, Baillie G, Ferguson C, Parton R, Wolvetang E, Roost M, Chuva de Sousa Lopes S, Little M
    Nature, 2015;526(7574):564-8.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Not Specified
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

CTS019 1
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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Isotype Controls
Description Application Cat# Citations Images  

Normal Sheep IgG Control

Ctrl 5-001-A 9
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Secondary Antibodies
Description Application Cat# Citations Images  

Sheep IgG HRP-conjugated Antibody

WB, Simple Western HAF016 14  
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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL010 3
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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL011 5
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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL012 2
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Sheep IgG (H+L) PE-conjugated Antibody

Flow F0126  
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Donkey Anti-Sheep IgG Biotinylated Antibody

WB BAF016 2
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Sheep IgG (H+L) APC-conjugated Antibody

Flow F0127  
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Sheep IgG VisUCyte HRP Polymer Antibody

IHC VC006  
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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

Flow F0125 1
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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

Flow F0128
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